RU2002133236A

RU2002133236A - NEW PHARMACEUTICAL COMPOSITION

Info

Publication number: RU2002133236A
Application number: RU2002133236/15A
Authority: RU
Inventors: Аполон ПАПАДИМИТРИОУ
Original assignee: Ф.Хоффманн-Ля Рош Аг
Priority date: 2000-05-15
Filing date: 2001-05-08
Publication date: 2004-06-27

Claims

1. A liquid pharmaceutical composition comprising an erythropoietin protein, a multiply charged inorganic anion in a pharmaceutically acceptable buffer, suitable for maintaining the pH of the solution in the range of about 5.5 to about 7.0, and optionally one or more pharmaceutically acceptable excipients.

2. The composition according to claim 1, which is an aqueous solution.

3. The composition according to claims 1 and 2, which is an isotonic solution.

4. The composition according to claims 1 to 3, where the anion is a multiply charged anion of a strong inorganic acid.

5. The composition according to claims 1 to 4, where the anion is selected from the group comprising sulfate, citrate or phosphate.

6. The composition according to claims 1-5, where the anion is sulfate.

7. The composition according to claim 6, containing from 10 to 200 mmol. / L sulfate.

8. The composition according to claims 1 to 7, where the pH is from 5.8 to 6.7.

9. The composition of claim 8, where the pH value is from 6.0 to 6.5.

10. The composition according to claim 9, where the pH value is approximately 6.2.

11. The composition according to claims 1-10, where the buffer is selected from the group comprising phosphate buffer or arginine / H ₂ SO ₄ / Na ₂ SO ₄ buffer.

12. The composition according to claim 11, where the buffer is a 10-50 millimolar phosphate buffer.

13. The composition according to claims 1-12, where the composition contains one or more pharmaceutically acceptable excipients.

14. The composition of claim 13, wherein the one or more pharmaceutically acceptable excipients is selected from the group consisting of pharmaceutically acceptable salts, diluents, solvents, and preservatives.

15. The composition according to claims 13 and 14, wherein the pharmaceutically acceptable excipients are selected from the group consisting of tonicity agents, polyols, antioxidants and nonionic surfactants.

16. The composition according to claims 13-15, wherein the pharmaceutically acceptable excipient is a polyol.

17. The composition according to clause 16, where the polyol is selected from the group comprising mannitol, sorbitol, glycerin, trehalose and sucrose.

18. The composition of claim 17, wherein the polyol is mannitol.

19. The composition according to PP.13-18, containing an antioxidant.

20. The composition according to claim 19, where the antioxidant is methionine.

21. The composition according to PP.13-20, containing up to 1 mmol / l CaCl ₂ .

22. The composition according to claims 15-22, wherein the nonionic surfactant is Polysorbate 80, Polysorbate 20 or Pluronic F68.

23. The composition according to item 22, where the nonionic surfactant is a pluronic F68.

24. The composition according to item 22 or 23, where the composition contains up to 1% (wt./about.) Non-ionic surfactants.

25. The composition according to any one of paragraphs.22-24, where the composition contains up to 0.1% (wt./about.) Non-ionic surfactants.

26. The composition of claims 1 to 25, wherein the erythropoietin protein is human erythropoietin.

27. The composition of claim 26, wherein the erythropoietin protein is expressed by endogenous gene activation.

28. The composition according to claims 26-27, wherein the erythropoietin protein has the amino acid sequence shown in SEQ ID NO: 1 or in SEQ ID NO: 2

29. The composition according to claims 26-28, wherein the protein has a human erythropoietin sequence modified by adding from 1 to 6 glycosylation sites.

30. The composition according to claims 26-28, wherein the erythropoietin protein has a human erythropoietin sequence modified by a modification selected from the group consisting of

Asn ³⁰ , Thr ³² ,

Asn ⁵¹ , Thr ⁵³ ,

Asn ⁵⁷ , Thr ⁵⁹ ,

Asn ⁶⁹ ,

Asn ⁶⁹ , Thr ⁷¹ ,

Ser ⁶⁸ , Asn ⁶⁹ Thr ³² ,

Val ⁸⁷ Asn ⁸⁸ , Thr ⁹⁰ ,

Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ ,

Ser ⁸⁷ Asn ⁸⁸ Gly ⁸⁹ Thr ⁹⁰ ,

Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ Thr ⁹² ,

Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ Ala ¹⁶² ,

Asn ⁶⁹ Thr ⁷¹ Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ ,

Asn ³⁰ Thr ³² Val ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ ,

Asn ⁸⁹ Ile ⁹⁰ Thr ⁹¹ ,

Ser87Asn ⁸⁹ Ile ⁹⁰ Thr ⁹¹ ,

Asn ¹³⁶ Thr ¹³⁸ ,

Asn ¹³⁸ Thr ¹⁴⁰ ,

Thr ¹²⁵ and

Pro ¹²⁴ Thr ^125.

31. The composition according to claims 26-30, wherein the sequence of human erythropoietin is modified by rearrangement of at least one glycosylation site.

32. The composition according to p, where the rearrangement includes a deletion of any of the N-linked glycosylation sites of human erythropoietin and the addition of an N-linked glycosylation site at position 88 of the sequence of human erythropoietin.

33. The composition according to p, where the glycoprotein has a human erythropoietin sequence modified by a modification selected from the group including

Gln ²⁴ Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ ,

Gln ³⁸ Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ and

Gln ⁸³ Ser ⁸⁷ Asn ⁸⁸ Thr ⁹⁰ .

34. The composition according to claims 26-33, wherein the erythropoietin protein indicated in any one of the paragraphs is pegylated.

35. The composition according to clause 34, where the erythropoietin protein is a conjugate, where the conjugate comprises an erythropoietin glycoprotein having at least one free amino group and having in vivo biological activity that causes an increase in the production of reticulocytes and erythrocytes by bone marrow cells, and selected from the group including human erythropoietin and its analogues, which have a human erythropoietin sequence modified by adding from 1 to 6 glycosylation sites or rearrangement by of at least one glycosylation site, this glycoprotein is covalently linked to the “n” poly (ethylene glycol) groups of the formula —CO— (CH ₂ ) _x - (OCH ₂ CH ₂ ) _m —OR, where the —CO radical of each poly (ethylene glycol) group forms an amide bond with one of these amino groups, R is (lower) alkyl, x = 2 or 3, m = 450-900, n = 1-3, and n and m are selected so that the molecular weight of the conjugate minus the molecular weight erythropoietin protein ranged from 20 to 100 kDa.

36. The composition according to clause 35, containing a protein erythropoietin of the formula

P- [NHCO- (CH ₂ ) _x - (OCH ₂ CH ₂ ) _m -OR] n (I)

where m, n, x and R are as defined above;

P is a glycoprotein residue without n amino groups (p), which (s) form an amide bond (s) with poly (ethylene glycol (s)) group (s).

37. The composition according to clause 36, where in the formula (I) x = 2, m = 650 - 750, n = 1 and R is methyl.

38. The composition according to clause 34, where the erythropoietin protein is a conjugate, and the conjugate contains an erythropoietin glycoprotein having at least one free amino group and having in vivo biological activity, causing an increase in the production of reticulocytes and erythrocytes by bone marrow cells, and which is selected from the group comprising human erythropoietin and its analogues having a primary structure of human erythropoietin modified by adding 1-6 glycosylation sites, where the covale glycoprotein It is clearly linked to one to three (lower) alkoxypoly (ethylene glycol) groups, each poly (ethylene glycol) group being covalently linked to the glycoprotein using a linker of the formula -C (O) -XSY with a C (O) fragment of the linker, as a result of which an amide bond with one of the amino groups, X is (CH ₂ ) _k - or -CH ₂ (O-CH ₂ -CH ₂ ) _k -, k = 1-10, Y is

or

the average molecular weight of each poly (ethylene glycol) fragment is from about 20 to about 40 kDa and the molecular weight of the conjugate is from about 51 to about 175 kDa.

39. The conjugate according to § 38 of the formula

where n = 1 to 3;

m = 450 - 900;

R is lower alkyl;

X is - (CH ₂ ) _k - or —CH ₂ (O — CH ₂ —CH ₂ ) _k -;

P denotes an erythropoietin glycoprotein residue without an amino group or groups forming an amide bond with X.

40. The conjugate according to § 39 of the formula

where H = 1 to 3;

m = 450 - 900;

R is lower alkyl;

X is - (CH ₂ ) _k - or —CH ₂ (O — CH ₂ —CH ₂ ) _k -;

41. The composition according to claims 1 to 40, containing from 10 to 10,000 μg of erythropoietin protein per millimeter.

42. The composition according to claims 1-41, containing from 10 to 10,000 μg of erythropoietin protein per millimeter, 10-200 mmol. / L sulfate and 10-50 mmol. / L phosphate, pH 6.0-6.5.

43. The composition according to item 42, containing up to 20 mm methionine, 1-5% (wt./about.) Polyol, up to 0.1% (wt./about.) Pluronic F68 and optionally up to 1 mm CaCl ₂ .

44. The composition according to item 42 or 43, containing 10-10000 μg of erythropoietin protein per millimeter, 40 mmol. / L sulfate, 10 mmol. / L phosphate, 3% (wt./about.) Mannitol, 10 mm methionine, 0 01% (w / v) pluronic F68, pH 6.2.

45. The composition according to claims 1-41, containing 10-10000 μg of erythropoietin protein per millimeter, 10-100 mol / L NaCl, 10-50 mmol / L phosphate, pH 6.0-7.0, optionally 1- 5% polyol.

46. The composition according to item 45, containing up to 20 mm methionine, up to 0.1% pluronic F68 and optionally 7.5 μmol / l CaCl ₂ .

47. The composition according to item 45 or 46, containing 10-10000 μg of erythropoietin protein per millimeter, 100 mmol./L NaCl, 10 mm methionine, 0.01% pluronic F68 and 10 mmol./L phosphate, pH 7.0.

48. The composition according to claims 1-41, containing 10-10000 μg of erythropoietin protein per millimeter, 10-50 mmol. / L of arginine, pH 6.0-6.5, 10-100 mmol. / L of sodium sulfate.

49. The composition according to p. 48, containing up to 20 mm methionine, up to 0.1% pluronic F68, optionally up to 1 μmol / l CaCl ₂ and optionally 1-5% polyol.

50. The composition according to p. 48 or 49, containing 10-10000 μg of erythropoietin protein per millimeter, 40 mmol. / L arginine, pH 6.2, 30 mmol. / L sodium sulfate, 3% mannitol, 10 mm methionine, 0, 01% pluronic F68, optionally 1 mmol / L CaCl ₂ .

51. The composition according to claims 1-41, containing 25-2500 μg / ml erythropoietin, and a) 10 mm sodium / potassium phosphate, 100 mm NaCl, pH 7.0 or b) 10 mm sodium phosphate, 120 mm sodium sulfate, pH 6.2 or c) 10 mM sodium phosphate, 40 mM sodium sulfate, 3% mannitol, pH 6.2 or g) 10 mM sodium phosphate, 40 mM sodium sulfate, 3% mannitol, 10 mM methionine, 0.01% pluronic F68, pH 6.2 or d) 40 mM arginine, 30 mM sodium sulfate, 3% mannitol, pH 6.2 or e) 40 mM arginine, 30 mM sodium sulfate, 3% mannitol, pH 6.2, 0, 01% pluronic F68, pH 6.2.

52. The composition according to claims 1-51, wherein the amount of erythropoietin is 50, 100, 400, 800 or 2500 μg / ml.

53. The composition according to paragraph 52, containing 10 mm sodium phosphate, 40 mm sodium sulfate, 3% mannitol, 10 mm methionine, 0.01% pluronic F68, pH 6.2.

54. The composition according to paragraph 52, containing 40 mm arginine, 30 mm sodium sulfate, 3% mannitol, 10 mm methionine, 0.01% pluronic F68, pH 6.2.

55. The composition according to claims 1-54, which is a lyophilisate or spray dried powder.

56. A method of producing a composition according to any one of claims 1 to 54, comprising mixing an erythropoietin protein with a solution containing anion containing many negative charges and optionally one or more pharmaceutically acceptable excipients.

57. The use of the composition according to any one of claims 1 to 54 for the preparation of medicines that can be used to treat and prevent diseases associated with anemia in patients with chronic renal failure (CRF), AIDS patients and / or for the treatment of cancer patients undergoing chemotherapy.

58. A method for the treatment and prevention of diseases accompanied by anemia in patients with chronic renal failure (CRF), AIDS patients and cancer patients undergoing chemotherapy, comprising the step of administering to the patient a composition according to any one of claims 1 to 54.

59. Device for local and systemic continuous administration of the composition according to any one of claims 1 to 54.

60. The invention as described above.